Dual Regulation of the mitotic exit network (MEN) by PP2A-Cdc55 phosphatase.

Exit from mitosis in budding yeast is triggered by activation of the key mitotic phosphatase Cdc14. At anaphase onset, the protease separase and Zds1 promote the downregulation of PP2A(Cdc55) phosphatase, which facilitates Cdk1-dependent phosphorylation of Net1 and provides the first wave of Cdc14 activity. Once Cdk1 activity starts to decline, the mitotic exit network (MEN) is activated to achieve full Cdc14 activation. Here we describe how the PP2A(Cdc55) phosphatase could act as a functional link between FEAR and MEN due to its action on Bfa1 and Mob1. We demonstrate that PP2A(Cdc55) regulates MEN activation by facilitating Cdc5- and Cdk1-dependent phosphorylation of Bfa1 and Mob1, respectively. Downregulation of PP2A(Cdc55) initiates MEN activity up to Cdc15 by Bfa1 inactivation. Surprisingly, the premature Bfa1 inactivation observed does not entail premature MEN activation, since an additional Cdk1-Clb2 inhibitory signal acting towards Dbf2-Mob1 activity restrains MEN activity until anaphase. In conclusion, we propose a clear picture of how PP2A(Cdc55) functions affect the regulation of various MEN components, contributing to mitotic exit.

Dual self-image technique for beam collimation

We propose an accurate technique for obtaining highly collimated beams, which also allows testing the collimation degree of a beam. It is based on comparing the period of two different self-images produced by a single diffraction grating. In this way, variations in the period of the diffraction grating do not affect to the measuring procedure. Self-images are acquired by two CMOS cameras and their periods are determined by fitting the variogram function of the self-images to a cosine function with polynomial envelopes. This way, loss of accuracy caused by imperfections of the measured self-images is avoided. As usual, collimation is obtained by displacing the collimation element with respect to the source along the optical axis. When the period of both self-images coincides, collimation is achieved. With this method neither a strict control of the period of the diffraction grating nor a transverse displacement, required in other techniques, are necessary. As an example, a LED considering paraxial approximation and point source illumination is collimated resulting a resolution in the divergence of the beam of σ φ = ± μrad.